What does calcein stain for?
Because Calcein AM does not require DNA binding, it stains all metabolically-active cells and can be used to measure metabolic activity in non-nucleated cells, such as platelets.
Does calcein stain dead cells?
Calcein is negatively charged and cell membrane- impermeant, and consequently is retained in the cytoplasm of viable cells with intact plasma membranes. Dead cells either do not stain with calcein due to lack of esterase activity, or fail to retain calcein in the cytoplasm due to compromised plasma membrane integrity.
What is calcein labeling?
In particular, calcein is used as a marker for identification of bone growth, which is indicated by a green color. Calcein green is a calcium chelator that adheres to regions of mineralizing bone thereby allowing localization of new bone.
How do you dissolve calcein am?
Calcein-AM labeling kit consists of lyophilized Calcein-AM and anhydrous DMSO. For reconstitution, bring the kit to room temperature and add 50 µL of DMSO to one vial of Calcein-AM dye until fully dissolved. Upon receipt, Calcein-AM should be stored at -20°C.
Is Calcein cytotoxic?
Calcein/AM is a fluorescent dye that localises intracellularly after esterase-dependent cellular trapping and which has shown cytotoxic activity against various established human tumour cell lines at relatively low concentrations.
How do you dilute calcein?
1. Remove one 50 µg calcein AM from the freezer and allow to warm to room temperature. 2. Add 50 µL high-quality, anhydrous DMSO to one vial calcein AM, yielding a 1 mM stock solution.
Is Calcein AM toxic?
Calcein, AM has been used for studies of enzymatic activity, cell membrane integrity, and long-term cell tracking due to its low cellular toxicity. It is important to note that for certain cell lines and experimental conditions, relatively high concentrations of Calcein, AM can be toxic to cells.
How do you take calcein am?
To stain at 1 µM, add 10 µL of the Calcein AM solution that had been further diluted 1:10 in PBS to 390 µL cells, forming a final volume of 400 µL. If different cell volumes were used, add Calcein AM appropriately. Mix by gently flicking the tubes. 10.5 Incubate cells at 37°C protected from light.
Is calcein soluble in water?
Calcein is a highly negatively charged, water-soluble fluorescein derivative. Its fluorescence is nearly independent of pH in the range between 6.5 and 12. The dye can be introduced into cells by microinjection and is useful for cell tracing and for the study of cell membrane integrity and fusion.
Is calcein AM toxic to cells?
How do you dissolve calcein?
Calcein AM, in contrast, is susceptible to hydrolysis. Therefore, it should be dissolved in organic solvents like DMSO with a concentration of approximately 10 mg/mL.
How do you make calcein solution?
Add 50 µL high-quality, anhydrous DMSO to one vial calcein AM to prepare a 1 mM stock solution. Add 10 µL of this stock solution to 190 µL DMSO to make a 50 µM working solution of calcein AM. Use this working solution within one day.
How do you dissolve Calcein am?
Is Calcein soluble in water?
Is Calcein a fluorophore?
Calcein-AM is a fluorogenic, cell-permeant fluorescent probe that indicates cellular health. When the acetoxymethyl ester is intact, this probe is nonfluorescent until acted upon by nonspecific esterases present within the healthy, live cells.